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近场光学显微术在生物大分子探测与功能研究中的应用 总被引:1,自引:0,他引:1
近场光学显微术是唯一一种具有单分子探测灵敏度,且在对生物样品研究时无损伤的一门新兴的高分辨光学显微术,它是根据近场光学理论在扫描探针显微术的基础上发展起来的。它突破了传统光学显微术衍射极限的限制,可在不损伤样品的情况下同时获得其形貌像和光学像,故在探测单个生物分子并研究其结构和功能以及分子间的相互作用等方面具有显著优势。本文主要介绍近几年来近场光学显微术在生物分子探测和功能研究,以及在分子生物学研究中的应用与进展。 相似文献
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量子点荧光探针是近几年发展起来的一种新型荧光标记物,拥有荧光染料及荧光蛋白所不能比拟的独特优势,已经在细胞功能研究及细胞表面和内部功能分子的探测、组织的成像和病灶的定位等方面得到了较为广泛的应用。本文对量子点的光学特性、生物化修饰及其在生物成像等方面的应用进展进行了较为详细的介绍,并展望了其应用发展。 相似文献
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近场光学显微镜具有nm量级的空间分辨率,量子点(quantum dots,QDs)荧光探针具有激发谱宽、发射谱线窄、荧光强度高、抗光漂白和稳定性高等优点,两者结合用于生物大分子的成像探测和识别具有广泛的应用前景。用近场光学显微镜对链霉亲和素偶联的QDs进行近场荧光激发,并对其荧光发射特性和光稳定性进行研究,结果表明:近场光学显微镜nm量级的空间分辨率,可以同时观察到了QDs的单体、二聚体和三聚体;QDs的荧光发射强度高,近场荧光像对比度好,单量子点的荧光半高宽达到25nm;对一定入射波长的单色激发光,QDs的近场荧光强度随着激发功率密度的增加线性增加,并很快趋于稳定。与传统的荧光染料如异硫氰酸荧光素相比,QDs的稳定性非常好,在激发功率密度为300W/cm2的近场辐射下,量子点的荧光强度超过6h基本保持不变,其抗光漂白能力远远高于普通荧光染料。 相似文献
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分子生物学自问世以来在各个领域都有着非常广泛的应用。随着分子生物学手段的不断发展,其在医学诊断和疾病治疗等领域的作用也获得医护人员的认可。分子生物学方法在血液病诊断和治疗方面的应用也尤为突出。本文综述了PCR,荧光原位杂交技术、基因芯片和二代测序等分子生物学手段在血液病中的应用,为血液病的诊断、治疗监测以及用药指导等方面的研究提供参考。 相似文献
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FISH技术在微生物生态学中的研究及进展 总被引:3,自引:0,他引:3
分子生物学技术在微生物生态学研究中具有灵敏、精确和快速的优势,但不能提供微生物的形态学、数量性状、空间分布等信息。荧光原位杂交技术结合了分子生物学的精确性和显微镜的可视性信息,可以在自然生境中监测和鉴定不同的微生物个体,尤其是对难培养和未被培养的微生物进行检测。荧光原位杂交技术被广泛用于微生物群落结构诊断和评价,现已成为微生物分子生态学研究中的热点技术。对荧光原位杂交技术的发展和在微生物分子生态学中的应用进行了综述,探讨了该技术应用中存在的问题和发展前景。 相似文献
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Alexa Fluor染料为罗丹明或香豆素的衍生物,是一种含有活性基团的有机荧光染料,具有激发光谱窄、发射光谱宽、量子产率高、光稳定性好,以及受温度、p H影响小等优点。近年来,Alexa Fluor染料作为荧光探针在细胞学和分子生物学研究领域取得了广泛应用,但对Alexa Fluor染料进行系统阐述的文章很少。因此,在综述Alexa Fluor染料的基本特征和优缺点的基础上,阐述其在细胞学和分子生物学研究中的应用,并对其应用和发展方向进行展望。 相似文献
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金纳米棒具有独特的光学性质、表面易修饰性、较低的生物毒性和良好的生物相容性,因而在成像、光热治疗和药物载带等方面具有极高的潜在应用价值.本文综述了典型的金纳米棒表面修饰方法及其在生物成像、光热治疗和药物治疗中的应用,重点阐述了通过金纳米棒同时实现肿瘤诊断和治疗相结合的研究进展. 相似文献
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活体动物体内光学成像技术的研究进展及其应用 总被引:2,自引:0,他引:2
活体动物体内光学成像是利用基因改构进行内源性成像试剂或外源性成像试剂标记细胞、蛋白或DNA,从而非侵入性地报告小动物体内的特定生物学事件的技术。活体成像可以直观灵敏地监测基因的表达模式、标记和示踪细胞、探讨蛋白间的相互作用,因而这一技术被广泛地用于分析基因的表达模式、评价基因治疗效果、评估肿瘤的发生和转移、监测移植器官等。简要综述了现有活体动物体内光学成像技术的基本原理、技术进展和相关应用。 相似文献
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Richard Cisek Leigh Spencer Nicole Prent Donatas Zigmantas George S. Espie Virginijus Barzda 《Photosynthesis research》2009,102(2-3):111-141
Emerging as well as the most frequently used optical microscopy techniques are reviewed and image contrast generation methods in a microscope are presented, focusing on the nonlinear contrasts such as harmonic generation and multiphoton excitation fluorescence. Nonlinear microscopy presents numerous advantages over linear microscopy techniques including improved deep tissue imaging, optical sectioning, and imaging of live unstained samples. Nonetheless, with the exception of multiphoton excitation fluorescence, nonlinear microscopy is in its infancy, lacking protocols, users and applications; hence, this review focuses on the potential of nonlinear microscopy for studying photosynthetic organisms. Examples of nonlinear microscopic imaging are presented including isolated light-harvesting antenna complexes from higher plants, starch granules, chloroplasts, unicellular alga Chlamydomonas reinhardtii, and cyanobacteria Leptolyngbya sp. and Anabaena sp. While focusing on nonlinear microscopy techniques, second and third harmonic generation and multiphoton excitation fluorescence microscopy, other emerging nonlinear imaging modalities are described and several linear optical microscopy techniques are reviewed in order to clearly describe their capabilities and to highlight the advantages of nonlinear microscopy. 相似文献
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Sarah J Erickson Jiajia Ge Andrea Sanchez Anuradha Godavarty 《Translational oncology》2010,3(1):16-22
Near-infrared (NIR) optical imaging is a noninvasive and nonionizing modality that is emerging as a diagnostic tool for breast cancer. The handheld optical devices developed to date using the NIR technology are predominantly developed for spectroscopic applications. A novel handheld probe-based optical imaging device has been recently developed toward area imaging and tomography applications. The three-dimensional (3D) tomographic imaging capabilities of the device have been demonstrated from previous fluorescence studies on tissue phantoms. In the current work, fluorescence imaging studies are performed on tissue phantoms, in vitro, and in vivo tissue models to demonstrate the fast two-dimensional (2D) surface imaging capabilities of this flexible handheld-based optical imaging device, toward clinical breast imaging studies. Preliminary experiments were performed using target(s) of varying volume (0.23 and 0.45 cm3) and depth (1–2 cm), using indocyanine green as the fluorescence contrast agent in liquid phantom, in vitro, and in vivo tissue models. The feasibility of fast 2D surface imaging (∼5 seconds) over large surface areas of 36 cm2 was demonstrated from various tissue models. The surface images could differentiate the target(s) from the background, allowing a rough estimate of the target''s location before extensive 3D tomographic analysis (future studies). 相似文献
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Longmire MR Ogawa M Hama Y Kosaka N Regino CA Choyke PL Kobayashi H 《Bioconjugate chemistry》2008,19(8):1735-1742
Optical imaging has the potential to improve the efficacy of surgical and endoscopic approaches to cancer treatment; however, the optimal type of fluorescent probe has not yet been established. It is well-known that rhodamine-core-derived fluorophores offer a combination of desirable properties such as good photostability, high extinction coefficient, and high fluorescence quantum yield. However, despite the ubiquitous use of rhodamine fluorophores for in vivo optical imaging, it remains to be determined if unique chemical properties among individual rhodamine core family members affect fluorophore parameters critical to in vivo optical imaging applications. These parameters include preserved fluorescence intensity in low pH environments, similar to that of the endolysosome; efficient fluorescence signal despite conformational changes to targeting proteins as may occur in harsh subcellular environments; persistence of fluorescence after cellular internalization; and sufficient signal-to-background ratios to permit the identification of fluorophore-targeted tumors. In the present study, we conjugated 4 common rhodamine-core based fluorescent dyes to a clinically feasible and quickly internalizing D-galactose receptor targeting reagent, galactosamine serum albumin (GmSA), and conducted a series of in vitro and in vivo experiments using a metastatic ovarian cancer mouse model to determine if differences in optical imaging properties exist among rhodamine fluorophores and if so, which rhodamine core possesses optimal characteristics for in vivo imaging applications. Herein, we demonstrate that the rhodamine-fluorophore, TAMRA, is the most robust of the 4 common rhodamine fluorophores for in vivo optical imaging of ovarian cancer metastases to the peritoneum. 相似文献
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Xianshao Zou Tingting Pan Lei Chen Weiwen Zhang 《Critical reviews in biotechnology》2017,37(6):723-738
Luminescence including fluorescence and phosphorescence sensors have been demonstrated to be important for studying cell metabolism, and diagnosing diseases and cancer. Various design principles have been employed for the development of sensors in different formats, such as organic molecules, polymers, polymeric hydrogels, and nanoparticles. The integration of the sensing with fluorescence imaging provides valuable tools for biomedical research and applications at not only bulk-cell level but also at single-cell level. In this article, we critically reviewed recent progresses on pH, oxygen, and dual pH and oxygen sensors specifically for their application in microbial cells. In addition, we focused not only on sensor materials with different chemical structures, but also on design and applications of sensors for better understanding cellular metabolism of microbial cells. Finally, we also provided an outlook for future materials design and key challenges in reaching broad applications in microbial cells. 相似文献
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Mertz J 《Nature methods》2011,8(10):811-819
A key requirement for performing three-dimensional (3D) imaging using optical microscopes is that they be capable of optical sectioning by distinguishing in-focus signal from out-of-focus background. Common techniques for fluorescence optical sectioning are confocal laser scanning microscopy and two-photon microscopy. But there is increasing interest in alternative optical sectioning techniques, particularly for applications involving high speeds, large fields of view or long-term imaging. In this Review, I examine two such techniques, based on planar illumination or structured illumination. The goal is to describe the advantages and disadvantages of these techniques. 相似文献
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Optical imaging can advance knowledge of cellular biology and disease at the molecular level in vitro and, more recently, in vivo. In vivo optical imaging has enabled real-time study to track cell movement, cell growth, and even some cell functions. Thus, it can be used in intact animals for disease detection, screening, diagnosis, drug development, and treatment evaluation. This review includes a brief introduction to fluorescence imaging, fluorescent probes, imaging devices, and in vivo applications in animal models. It also describes a quantitative fluorescence detection method with a reconstruction algorithm for determining the location of fluorophores in tissue and addresses future applications of in vivo fluorescence imaging. 相似文献